LOCUS       sequence-233593-        3435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNA targeting the 3'-untranslated region (UTR) of the gamma2 mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3435)
  AUTHORS   Li RW, Yu W, Christie S, Miralles CP, Bai J, Loturco JJ, De Blas AL
  TITLE     Disruption of postsynaptic GABA receptor clusters leads to decreased
            GABAergic innervation of pyramidal neurons.
  JOURNAL   J Neurochem. 2005 Nov;95(3):756-70. doi: 
            10.1111/j.1471-4159.2005.03426.x.
  PUBMED    16248887
REFERENCE   2  (bases 1 to 3435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1111/j.1471-4159.2005.03426"; journalName: "J Neurochem"; date: 
            "2005-11"; volume: "95"; issue: "3"; pages: "756-70"
COMMENT     Sequence Label: gamma2 UTR shRNA (shRNA4)
FEATURES             Location/Qualifiers
     source          1..3435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        627..939
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     858..878
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     polyA_signal    1013..1147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1037..1056)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1091..1110
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1245..1265)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1245..1263)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1284..1300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1284..1300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1297..1319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1308..1324
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1332..1338,1339..1356,1357..1362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1377..1398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1515..1532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1686..2274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1766..1785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2445..3236,3237..3305))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3068..3087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3306..3410)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg acagcctgca
      541 ggaatggccg gccatcttaa ttaagaaatt taaatatgtt taaacatagg gcccaatggc
      601 gcgccaaggc catctaggcc aagcttatcc gacgccgcca tctctaggcc cgcgccggcc
      661 ccctcgcaca gacttgtggg agaagctcgg ctactcccct gccccggtta atttgcatat
      721 aatatttcct agtaactata gaggcttaat gtgcgataaa agacagataa tctgttcttt
      781 ttaatactag ctacatttta catgataggc ttggatttct ataagagata caaatactaa
      841 attattattt taaaaaacag cacaaaagga aactcaccct aactgtaaag taattgtgtg
      901 ttttgagact ataaatatcc cttggagaaa agccttgttt ggtgagattc agcgaataag
      961 acccctacaa ggggtcttat acgctaatct cactttttct agataatacg tagatccaga
     1021 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1081 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1141 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1201 ggttttttaa ttcttgcggc cgcggcggta cccagctttt gttcccttta gtgagggtta
     1261 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1321 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     1381 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1441 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1501 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1561 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1621 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1861 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2101 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2281 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3421 cgaaaagtgc cacct
//
